Hazardous biochemical agents in animal husbandry indoor environments are known to promote the occurrence of various illnesses among workers and animals. The relationship between endotoxin levels in dust collected from chicken farms and various immunological markers was investigated. Peripheral blood was obtained from 20 broiler chickens and 20 laying hens from four different chicken farms in Korea. Concentrations of total or respirable dust in the inside the chicken farm buildings were measured using a polyvinyl chloride membrane filter and mini volume sampler. Endotoxin levels in the dust were determined by the Limulus Amebocyte Lysate Kinetic method. Interferon-gamma production by peripheral blood mononuclear cells stimulated with concanavalin A was significantly lower in broilers or layers from the farms with higher endotoxin concentrations than the chickens from the farms with lower endotoxin levels. An opposite pattern was observed for plasma cortisol concentrations with higher cortisol levels found in chickens from the farms with higher endotoxin levels. When peripheral lymphocytes were examined, the percentage of CD3-Ia+ B cells was lower in layers from farms with higher endotoxin levels than those from locations with lower endotoxin levels. Overall, these results suggest a probable negative association between dust endotoxin levels and cell-mediated immunity in chickens.
Animal Husbandry ; Animals ; Biomarkers/blood ; Chickens/*immunology ; Dust/*analysis ; Endotoxins/*analysis ; *Housing, Animal ; *Immunity, Cellular

Pillows are known to contain significant levels of indoor allergens and endotoxin, that are of importance to house dust mite sensitized asthmatics. Buckwheat pillows are commonly used in Korea. We studied the levels of the house dust mite allergen, Der f 1, and endotoxin on new synthetic and buckwheat pillows and their accumulation over three months. Endotoxin levels were significantly higher on new buckwheat pillows compared to synthetic pillows; geometric mean levels (95% CI) were 60,950 EU/g (30,270-122,700) and 4,887 EU/g (2,570-9,311) respectively (p<0.001). No Der f 1 was detected on the new pillows. After three months Der f 1 levels were similar on buckwheat and synthetic pillows, geometric mean levels (95% CI) were 1.16 microgram/g (0.02-8.13) and 1.08 microgram/g (0.19-1.68) respectively. These results indicate that buckwheat pillows are a source of very high endotoxin levels that may be of relevance to asthma severity of atopic asthmatics.
Allergens/*immunology ; Animals ; Antigens, Dermatophagoides/*immunology ; Asthma/immunology ; *Bedding and Linens ; *Buckwheat ; Dermatophagoides farinae/immunology ; Endotoxins/*immunology ; Humans ; Korea ; Research Support, Non-U.S. Gov't

INTRODUCTIONThis study investigated the responses of plasma endotoxin and pro- and antiinflammatory cytokines during a 21-km road race in warm and humid conditions. The influence of carbohydrate-electrolyte (CE)-water (WA) drink mix ingested on leukocyte subset responses and the association between plasma lipopolysaccharide (LPS) concentration and fluid balance, exercise intensity, and body core temperature (Tc) were also studied.

MATERIALS AND METHODSThirty runners provided blood samples before and after the half-marathon for leukocyte, LPS and cytokine analyses. Tc was measured by the ingestible telemetric temperature sensor and fluid intake and split-times were recorded at 3 km intervals. Exercise intensity was determined by matching running speed and heart rate during the race with the corresponding speed-oxygen uptake relationship and heart rate measured in the laboratory 2 to 6 weeks before the race.

RESULTSPlasma LPS concentration increased from 1.9 +/- 1.9 pg/mL before, to 2.5 +/- 1.9 pg/mL after running (P <0.05). Peak plasma LPS concentration was 7.5 pg/mL. Plasma IL-1beta and TNF-concentration did not change significantly, whereas significant increases in IL-10 (50%), IL-1ra (23.2%) and IL-6 (65.2%) were observed after the race. No significant correlation between plasma LPS concentration and exercise intensity, hydration and Tc was observed.

CONCLUSIONLeukocyte subset responses were not related to the ratio of CE and water drink mix ingested. Running a half-marathon can induce mild endotoxaemia, which is not related to exercise intensity, fluid balance, and Tc responses. Mixing CE drink with water did not mitigate postexercise leukocytosis and lymphopenia.

Adult ; Beverages ; Cytokines ; immunology ; Endotoxins ; blood ; immunology ; Fluid Therapy ; Hot Temperature ; Humans ; Humidity ; Leukocytes ; immunology ; Lipopolysaccharides ; immunology ; Male ; Physical Exertion ; physiology ; Running ; physiology

OBJECTIVETo explore the mechanism of endotoxin tolerance (ET) through observing the expression of interleukin 1 receptor associated kinase-4 (IRAK-4) during endotoxin tolerance development in Kupffer cells (KCs).

METHODSIsolated KCs of Balb/c mouse were divided into two groups: the non-endotoxin tolerance (NET) group and the endotoxin tolerance (ET) group, which were pretreated with 10 ng/ml lipopolysaccharide (LPS) for 24 h. Then, the two groups were treated with 100 ng/ml LPS. The expressions of IRAK-4 gene and protein level were determined by RT-PCR and Western blot. The activities of NF-kappaB of KCs and the TNFalpha level were estimated by ELISA at 0 h, 1 h, 3 h, 6 h and 12 h after LPS stimulation.

RESULTSThe ultimate level of IRAK-4, the activities of NF-kappaB and the TNFalpha level were evidently lower in the ET group than those in the NET group (t = 12.4, 17.4 and 138.9 respectively, P<0.01).

CONCLUSIONSPretreatment with LPS on KCs could induce endotoxin tolerance of KCs and inhibition of IRAK-4 expression may be one of the reasons for its development.

Animals ; Cells, Cultured ; Endotoxins ; immunology ; Immune Tolerance ; Interleukin-1 Receptor-Associated Kinases ; biosynthesis ; genetics ; Kupffer Cells ; cytology ; immunology ; metabolism ; Lipopolysaccharides ; immunology ; Male ; Mice ; Mice, Inbred BALB C

A series of pathophysiological changes in lymph circulation system occur after severe burns. We try to elucidate the importance through summarizing our experiments on some of the changes in lymph circulation based on rat and goat lymphatic fistula model since 1998. The lymphatic contraction frequency decreased while the lymph flow speed increased during burn shock stage. Contents of several key inflammatory factors, such as TNF-alpha, IL-6, IL-8, IL-10, IL-18, and HMGB-1, were increased in lymph or lymph nodes, and they were higher than those in blood and liver. The protein concentration increased in lymph while decreased in plasma. The endotoxin was translocated to lymph earlier than to blood, therefore, the number of E. coli or the number of endotoxin translocated via lymph route were more than those via blood. The bacteria and endotoxin of pseudomonas aeruginosa could invade through local lymphatic route from infected burn wound. Th2 shift from Th1/Th2 occurred in lymph and the ratio of CD4+/CD8+ T lymph cells decreased in lymph nodes after burns, denoting local immunosuppression. The apoptosis of lymphocytes in lymph organ might contribute to this immunosuppression.
Animals ; Burns ; immunology ; metabolism ; microbiology ; CD4-CD8 Ratio ; Disease Models, Animal ; Endotoxins ; adverse effects ; Goats ; Inflammation Mediators ; immunology ; Lymphatic System ; metabolism ; Rats ; Th2 Cells ; immunology

OBJECTIVETo investigate the enhancing effect of Chinese medicine-Xuebijing injection on lipopolysaccharide (LPS) -induced apoptosis of CD4+ CD25+ regulatory T cells (Tregs) and polarization of helper T cells (Th).

METHODSCD4+ CD25+ Tregs collected from rat spleen in vitro by immunomagnetic beads assay were divided into the control group, anti-CD3/CD28 group, anti-CD3/CD28 + LPS group, anti-CD3/CD28 + "Xuebijing injection" group and anti-CD3/CD28 + LPS + "Xuebijing injection" group. Tregs apoptosis rate and expression of winged helix transcription factor (Foxp3) in Tregs were detected by flow cytometry on 3rd post culture day. CD4+ CD25- T cells were co-cultured with CD4+ CD25- Tregs (1:1) for 68 hours with canavalin A stimulation. Interferon gamma (gamma-IFN), interleukin (IL)-4 and IL-17 in supernatants, which respectively was secreted by Th1, Th2 and Th17, were measured by ELISA.

RESULTSTregs apoptosis rate of anti-CD3/CD28 + LPS + "Xuebijing injection" group (45.1 +/- 2.7%) was significantly higher than that of anti-CD3/CD28 + LPS group (29.4 +/- 1.6%, P < 0.01). Meanwhile, Foxp3 expressions in Tregs in above 2 groups were 95 +/- 9 and 140 +/- 18 respectively, showing statistically significant difference between them (P < 0.01). Gamma-IFN levels secreted in anti-CD3/CD28 + LPS + "Xuebijing injection" group were significantly higher than those in anti-CD3/CD28 + LPS group (P < 0.01), while IL-4 levels had an opposite tendency compared with gamma-IFN (P < 0.05), resulting in a marked increase in the ra- tio of gamma-IFN/IL-4 in anti-CD3/CD28 + LPS + "Xuebijing injection" group (P < 0.01). In anti-CD3/ CD28 + "Xuebijing injection" group, IL-17 secretion levels were significantly decreased compared with anti-CD3/CD28 group (P < 0.05).

CONCLUSIONSActivation of CD4+ CD25+ Tregs induced by LPS may mediate Th1 shift to Th2 response. "Xuebijing injection" can effectively regulate immune function of T cells, increase the LPS-induced apoptosis of CD4+ CD25+ Tregs as well as enhance the polarization of Th2 to Th1, thereby abating the suppressive state of cell-mediated immunity.

Animals ; Apoptosis ; Drugs, Chinese Herbal ; pharmacology ; Endotoxins ; Lipopolysaccharides ; Male ; Rats ; Rats, Wistar ; T-Lymphocytes, Helper-Inducer ; drug effects ; immunology ; T-Lymphocytes, Regulatory ; drug effects ; immunology

OBJECTIVEBy using RAW 264.7 macrophage cell line, we studied the dose-effect relationship of endotoxin induced RAW 264.7 cells to release TNF-alpha, and then detected the content of endotoxin in 8 kinds of injections, so that we can investigate the feasibility and the interference factors of the novel test.

METHODBy using endotoxin of different concentrations to induce RAW 264. 7 cells to release TNF-a, we drew the curve of dose-effect relationship between endotoxin and generated TNF-alpha. Then we detected the content of TNF-alpha in yuxingcao, shuanghuanglian, qingkailing, gegensu, xiangdan, qianrongmei and jiangxianmei injections and shuanghuanglian powder injection, and calculated their content of endotoxin.

RESULTThe endotoxin could induce the cells to release TNF-alpha in a good dose-dependent manner, even at a very low concentration. In the range of maximum available dilution multiple, the content of endotoxin in the rest 7 kinds of injections was less than 1.0 EU x mL(-1) except qingkailing injection of two batch.

CONCLUSIONCytokine revulsion has the advantage of wide detection range, high sensitivity, simple operation, and the detected endotoxin is of bioactivity. This method provides another technical mean for pyrogen test of injections.

Animals ; Biological Assay ; methods ; Cell Line ; Drug Contamination ; Drugs, Chinese Herbal ; analysis ; Endotoxins ; adverse effects ; analysis ; Macrophages ; drug effects ; immunology ; Mice ; Tumor Necrosis Factor-alpha ; analysis ; immunology

OBJECTIVETo explore the changes of cytokines including TNFalpha, TGFbeta1 and nitrogen monoxide, and endotoxin in the serum of chronic severe hepatitis after the treatment of ALSS, and to evaluate further the value of ALSS in the treatment of chronic severe hepatitis.

METHODSForty two patients were screened. The changes of TNFalpha, TGFbeta1, nitrogen monoxide and endotoxin were detected respectively. The relationship between the cytokines and the severity and prognosis were further analyzed.

RESULTSALSS was effective to decrease the serum concentration of cytokines. TNFalpha dropped from (481.57+/-229.33) pg/ml to (156.46+/-78.12) pg/ml (P < 0.05). TGFbeta1 from (44.09+/-31.73) ng/ml to (27.77+/-23.28) ng/ml (P < 0.01), endotoxin from (1.05+/-0.37) Eu/ml to (0.28+/-0.22) Eu/ml (P < 0.001). NO from (71.15+/-33.09) micromol/L to (58.11+/-29.30) micromol/L (P < 0.001). Before the therapy endotoxin was related with TNFalpha and total bilirubin, while after the therapy, NO was related with protime and aminonemia.

CONCLUSIONHigh level of endotoxin and nitrogen monoxide in serum plays an important role in hepatocyte damage of chronic severe hepatitis. The changes of serum endotoxin TNFalpha, TGFbeta1 and nitrogen monoxide level in patients with chronic severe hepatitis can be used to judge the severity and prognosis of severe hepatitis. ALSS is a reliable hepatic support device for chronic severe hepatitis

Adult ; Aged ; Cytokines ; blood ; Endotoxins ; blood ; Female ; Hepatitis, Chronic ; blood ; immunology ; therapy ; Humans ; Liver, Artificial ; Male ; Middle Aged ; Nitric Oxide ; blood ; Prognosis ; Transforming Growth Factor beta ; blood

This study was aimed to prepare the polypeptide of N-terminal heparin-binding domain of fibronectin(rhFNHN-29 polypeptide) with pichia expression system, to detect biological activity of recombinant polypeptide and investigate its effect on disseminated intravascular coagulation (DIC) in rats. The sequence of N-terminal heparin-binding domain of fibronectin was amplified from FNcDNA by PCR. The aim gene was cloned into T vector for selection. Then it was cloned into pAo815SM and pPIC9K vectors.Lined pPIC9K vectors were transformed into GS115 Pichia cells so as to express the aim polypeptide in Pichia expression system. The fermentation liquid were precipitated by 80% ammonium sulfate, and the further dissolved sediment were purified using S-100 column and SP column. Its activity of binding with heparin were detected by Western-blot. The established DIC rats (40 rats) were randomly divided into two groups. One group was treated with rhFNHN-29 polypeptide, and the other was treated with normal saline. The rats in the former group were injected with rhFNHN-29 polypeptide (10 mg/kg) through tail vein at 0.5 hour before, 2 hours and 4 hours after injection of LPS respectively. The rats in latter group were injected with equal volume saline. In addition, 20 normal rats injected with normal saline were as normal controls. 500 microl blood was taken from the rat vein, at 6 hours after the injection of LPS. White blood cell (WBC), hemoglobin (Hb) and platelets were tested from 50 microl blood. The rest 450 microl blood was used to isolate plasma for detecting TNFa level and coagulogram. The rats were killed at 24 hours after injection with LPS. Their livers, lungs, hearts, kidneys, and brain tissues were taken for histopathologic examination. The results showed that the aim polypeptide was successfully expressed in Pichia expression system. The expression level reached approximately 30 mg/L. The polypeptide had activity of binding with heparin antibody. In the experiment study of polypeptide effect on DIC in rats, the plasma TNFa level in polypeptide-treated group was lower than that in saline control group, the hemogram, coagulogram and histopathology were more obviously improved in polypeptide-treated group as compared with saline control group. It is concluded that the rhFNHN-29 polypeptide is successfully prepared, this polypeptide can antagonize DIC induced by endotoxin in rats.
Animals ; Disseminated Intravascular Coagulation ; therapy ; Endotoxins ; Female ; Fibronectins ; genetics ; immunology ; therapeutic use ; Heparin ; metabolism ; Male ; Peptides ; genetics ; therapeutic use ; Pichia ; metabolism ; Rats ; Rats, Sprague-Dawley

PURPOSE: Cockroach (CR) is an important inhalant allergen and can induce allergic asthma. However, the mechanism by which CR induces airway allergic inflammation and the role of endotoxin in CR extract are not clearly understood in regards to the development of airway inflammation. In this study, we evaluated whether endotoxin is essential to the development of CR induced airway allergic inflammation in mice. MATERIALS AND METHODS: Airway allergic inflammation was induced by intranasal administration of either CR extract, CR with additional endotoxin, or endotoxin depleted CR extract, respectively, in BALB/c wild type mice. CR induced inflammation was also evaluated with toll like receptor-4 (TLR-4) mutant (C3H/HeJ) and wild type (C3H/HeN) mice. RESULTS: Intranasal administration of CR extracts significantly induced airway hyperresponsiveness (AHR), eosinophilic and neutrophilic airway inflammation, as well as goblet cell hyperplasia in a dose-dependent manner. The addition of endotoxin along with CR allergen attenuated eosinophilic inflammation, interleukin (IL)-13 level, and goblet cell hyperplasia of respiratory epithelium; however, it did not affect the development of AHR. Endotoxin depletion in CR extract did not attenuate eosinophilic inflammation and lymphocytosis in BAL fluid, AHR and IL-13 expression in the lungs compared to CR alone. The attenuation of AHR, eosinophilic inflammation, and goblet cell hyperplasia induced by CR extract alone was not different between TLR-4 mutant and the wild type mice. In addition, heat inactivated CR extract administration induced attenuated AHR and eosinophilic inflammation. CONCLUSION: Endotoxin in CR extracts may not be essential to the development of airway inflammation.
Allergens/*immunology ; Animals ; Asthma/*chemically induced/*immunology/metabolism ; Cockroaches/*immunology ; Endotoxins/*immunology ; Enzyme-Linked Immunosorbent Assay ; Female ; Inflammation/*chemically induced/*immunology/metabolism ; Interferon-gamma/metabolism ; Interleukin-13/metabolism ; Interleukin-5/metabolism ; Mice ; Mice, Inbred BALB C ; Respiratory Hypersensitivity/chemically induced/*immunology